The Absence of PTEN in Breast Cancer Is a Driver of MLN4924 Resistance

Abstract

Background: Numerous studies have indicated that the neddylation pathway is closely associated with tumor development. MLN4924 (Pevonedistat), an inhibitor of the NEDD8-activating E1 enzyme, is considered a promising chemotherapeutic agent. Recently, we demonstrated that neddylation of the tumor suppressor PTEN occurs under high glucose conditions and promotes breast cancer development. It has been shown, however, that PTEN protein levels are reduced by 30–40% in breast cancer. Whether this PTEN deficiency affects the anti-tumor function of MLN4924 is unknown.Methods: In the present study, cell counting kit-8 and colony formation assays were used to detect cell proliferation, and a transwell system was used to quantify cell migration. A tumor growth assay was performed in BALB/c nude mice. The subcellular location of PTEN was detected by fluorescence microscopy. The CpG island of the UBA3 gene was predicted by the Database of CpG Islands and UCSC database. Western blotting and qRT-PCR were used to measure the expression of indicated proteins. The Human Protein Atlas database, the Cancer Genome Atlas and Gene Expression Omnibus datasets were used to validate the expression levels of UBA3 in breast cancer.Results: Our data show that the anti-tumor efficacy of MLN4924 in breast cancer cells was markedly reduced with the deletion of PTEN. PI3K/Akt signaling pathway activity correlated positively with UBA3 expression. Pathway activity correlated negatively with NEDP1 expression in PTEN-positive breast cancer patients, but not in PTEN-negative patients. We also demonstrate that high glucose conditions upregulate UBA3 mRNA by inhibiting UBA3 promoter methylation, and this upregulation results in the overactivation of PTEN neddylation in breast cancer cells.Conclusion: These data suggest a mechanism by which high glucose activates neddylation. PTEN is critical, if not indispensable, for MLN4924 suppression of tumor growth; PTEN status thus may help to identify MLN4924-responsive breast cancer patients.

Neddylation inactivation affects cell cycle and apoptosis in sheep follicular granulosa cells

“…MLN4924 overactivated the AKT pathway in a feedback manner to promote the survival signaling in sheep follicular GCs, which could also lead to MLN4924 resistance to apoptosis; similar findings have been confirmed in other studies (Chen et al, 2018;Fujita et al, 2006;Mao et al, 2019;Shi et al, 2006). A highthroughput RNA-Seq study found that FOXO1 was upregulated upon MLN4924 treatment (Du et al, 2021), and our results also found an upregulation of FOXO1 that could promote sheep follicular GCs apoptosis. However, the expression of phosphorylated FOXO3a at Thr32 was downregulated by MLN4924 treatment, which could be due to the over-activation of AKT inhibiting FOXO3a transactivation (Wang et al, 2017).…”

The Cellular and Developmental Roles of Cullins, Neddylation, and the COP9 Signalosome in Dictyostelium discoideum

“…In addition, counting factor-associated protein D (CfaD), which is a secreted quorum sensing protein that modulates cell proliferation, was detected in the FbxD interactome ( Figure 6 ; Bakthavatsalam et al, 2008 ; Sheikh et al, 2015 ). These findings indicate a conserved role for neddylation in regulating cell proliferation and support recent work linking neddylation to the proliferation of cancer cells ( Du et al, 2021 ; Zhang et al, 2021 ). However, whether MLN4924 specifically affects CRLs in D. discoideum remains to be determined since non-cullin targets of neddylation have been reported in several organisms including S. pombe , A. thaliana , and Trypanosoma brucei ( Girdwood et al, 2012 ; Enchev et al, 2015 ; Mergner et al, 2015 ; Liao et al, 2017 ).…”

SUMOylation and NEDDylation in Primary and Metastatic Cancers to Bone

“…Moreover, high level NEDDylation of PTEN in BC patients correlated with poor overall survival, indicating that PTEN NEDDylation might function as a prognostic marker ( Xie et al, 2021b ). Very recently, it has been reported that high glucose concentrations trigger PTEN NEDDylation and nuclear import and promote tumor development ( Du et al, 2021 ).…”

“…Another study reported about the resistance to MLN4924, a selective inhibitor of NAE1 and thus of the NEDDylation modification ( Soucy et al, 2009 ), treatment in cancer patients with complete loss of PTEN, indicating that PTEN is necessary for MLN4924 suppression of tumor growth and that PTEN status is useful to discriminate MLN4924-responsive BC patients ( Du et al, 2021 ).…”